The effect of surface probe density on DNA hybridization

AUTOR(ES)

Peterson, Alexander W.

FONTE

Oxford University Press

RESUMO

The hybridization of complementary strands of DNA is the underlying principle of all microarray-based techniques for the analysis of DNA variation. In this paper, we study how probe immobilization at surfaces, specifically probe density, influences the kinetics of target capture using surface plasmon resonance (SPR) spectroscopy, an in situ label-free optical method. Probe density is controlled by varying immobilization conditions, including solution ionic strength, interfacial electrostatic potential and whether duplex or single stranded oligonucleotides are used. Independent of which probe immobilization strategy is used, we find that DNA films of equal probe density exhibit reproducible efficiencies and reproducible kinetics for probe/target hybridization. However, hybridization depends strongly on probe density in both the efficiency of duplex formation and the kinetics of target capture. We propose that probe density effects may account for the observed variation in target-capture rates, which have previously been attributed to thermodynamic effects.

Documentos Relacionados

• Oligodeoxyribonucleotide probe accessibility on a three-dimensional DNA microarray surface and the effect of hybridization time on the accuracy of expression ratios
• DNA hybridization on microparticles: determining capture-probe density and equilibrium dissociation constants.
• DNA hybridization probe for the Pseudomonas fluorescens group.
• DNA hybridization probe for clinical diagnosis of Entamoeba histolytica.
• Effect of ligand binding on the buoyant density of DNA in Nycodenz gradients.